An efficient method for purifying high quality RNA from wheat pistils

Manickavelu, Alagu; Kambara, Kumiko; Mishina, Kohei; Koba, Takato

doi:10.1016/j.colsurfb.2006.10.024

Cited by 36 publications

(33 citation statements)

References 14 publications

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“…Así mismo, la extracción de RNA de tejidos de plantas recalcitrantes conlleva una serie de problemas por la presencia de pectinas, taninos (ME., 1992), pigmentos (Kim y Hamada, 2005), polisacáridos (Manickavelu et al, 2007), poliquinonas (Meisel et al, 2005) y otros metabolitos secundarios (de Keukeleire et al, 2006). Todos estos componentes vegetales pueden interactuar con los ácidos nucleicos, concretamente con el RNA formando complejos insolubles.…”

Section: Caracterización Del Gen Ppo1-tcunclassified

Identificación de un gen codificante de polifenol oxidasa (PPO) en Theobroma cacao L. (cacao) de Ecuador

Carriel

Acosta-Farías

Huebla-Concha

et al. 2017

CyT

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En Ecuador, las plantaciones de cacao presentan bajos promedios de producción debido a la diversidad de patógenos, especialmente a la infección por Moniliophthora roreri (monilia). Se cree que existe una relación entre el ataque del hongo y el aumento de los niveles de expresión de genes codificantes de polifenol oxidasas (PPOs) como mecanismo de defensa ante patógenos y herbívoros en diferentes plantas. Para la identificación de genes que codifican para PPOs, se seleccionaron hojas de cacao Nacional, provenientes de plantas resistentes y susceptibles a monilia, ubicadas en la Finca Experimental La Represa, propiedad de la Universidad Técnica Estatal de Quevedo. Se afinó un protocolo de extracción de ARN total de alta calidad para hojas de cacao recalcitrantes. Después de su retrotranscripción a ADNc, se realizaron ensayos de amplificación por PCR con diferentes primers, diseñados a partir de secuencias conservadas de PPOs. Los productos de amplificación permitieron la identificación de un gen de 961 pb, similar a un gen que codifica para la PPO predictiva de Theobroma cacao depositada en NCBI (XP_017978715.1) La identificación de este gen, es fundamental para evaluar a futuro los niveles de expresión y cuantificación en diferentes estados de desarrollo del fruto. Dicha cuantificación permitirá proponer herramientas de control para monilia y construir las bases para el mejoramiento genético del cacao Nacional.

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Section: Caracterización Del Gen Ppo1-tcunclassified

Identificación de un gen codificante de polifenol oxidasa (PPO) en Theobroma cacao L. (cacao) de Ecuador

Carriel

Acosta-Farías

Huebla-Concha

et al. 2017

CyT

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“…First-strand cDNA and double-strand cDNA were synthesized according to the protocols described by Manickavelu et al (2007). Some steps were modified as necessary.…”

Section: Double-strand Cdna Synthesismentioning

confidence: 99%

Analysis of gene expression patterns and levels in maize hybrids and their parents

Hs¹,

Li²,

Xh³

et al. 2015

Genet. Mol. Res.

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ABSTRACT. Heterosis has greatly contributed to conventional plant breeding and is widely used to increase crop plant productivity. However, although some studies have explored the mechanisms of heterosis at the genomic and transcriptome level, these mechanisms still remain unclear. The growth and development of maize seedlings and immature embryos have an important impact on subsequent production. This study investigated differentially expressed genes (DEGs) between parents and reciprocal hybrids in the seedling leaves, roots, and immature embryo 15 days after pollination using amplified fragment length polymorphism (AFLP)-based transcript profiling (cDNA-AFLP). We isolated 180, 170, and 108 genes from the leaves, roots, and immature embryos, respectively, that were differentially expressed between hybrids and parents. Sequencing and functional analysis revealed that 107 transcriptderived fragments in the roots and leaves and 90 in the immature embryos were involved in known functions, whereas many DEGs had roles in plant growth and development, photosynthesis, signal transduction, and seed germination. Quantitative reverse-transcription polymerase chain reaction analysis of relative expression levels between reciprocal hybrids and both parental genotypes of selected genes produced results that 15400 H.S. Nie et al. ©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 14 (4): 15399-15411 (2015) were consistent with cDNA-AFLP. We validated the expression patterns of 15 selected genes related to heterosis formation and revealed that most showed non-additive expression in one or both hybrids, including dominant, underdominant, and overdominant expression. This indicates that gene-regulatory interactions among parental alleles play an important role in heterosis during the early developmental stages of maize.

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“…The obtained DNA was dissolved in Tris-EDTA buffer and kept at -20°C. Total RNA was isolated from young spikes according to methods described in Manickavelu et al (2007). The total RNA was dissolved in RNase-free double-distilled water, and kept at -70°C.…”

Section: Dna and Rna Isolationmentioning

confidence: 99%

Cloning and characterization of thioredoxin h in the three-pistil line of common wheat

Yang¹,

Peng²,

Wu³

et al. 2013

Genet. Mol. Res.

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ABSTRACT. Thioredoxin h (Trxh) is a ubiquitous protein that reduces disulfides in target proteins, and is itself reduced by NADPHthioredoxin reductase. In the current study, the complementary DNA sequence and the genomic sequence of the three-pistil (TP) line of common wheat (Triticum aestivum L.) were obtained from spikes through reverse transcription-polymerase chain reaction (RT-PCR) and touchdown-PCR. Sequence alignment of amino acids of TPTrxh then allowed for predictions of its physicochemical properties, secondary structures, tertiary structures, and functional domains. Furthermore, the TPTrxh gene was overexpressed in Escherichia coli and its activity was demonstrated using a dithiothreitol-dependent insulin assay. The expression patterns of TPTrxh were analyzed through real-time RT-PCR in different tissues and across different developmental stages of young spikes. The complementary DNA of TPTrxh was found to be 411 bp in length, encoding 118 amino acids. Its genomic sequence was determined to be 2632 bp, possessing 3 exons and 2 introns. Functional domain analysis indicated that TPTrxh contained a WCGPC motif located at the end of the second β-fold and on the initial side of the second α-helix. The TPTrxh protein reduces intramolecular and intermolecular Cloning and characterization of thioredoxin h in TP lines disulfide bridges in target proteins. Young spikes contain higher levels of TPTrxh transcripts than do stems and leaves. The transcript levels in the young spikes (2-5 mm in length) of the Chinese Spring TP line increased 2.84-fold relative to those of young spikes (2-5 mm in length) of the Chinese Spring line. These data provide a basis for future research into the function of Trxh, and offer further insight into the molecular mechanism of the TP mutation in wheat.

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An efficient method for purifying high quality RNA from wheat pistils

Cited by 36 publications

References 14 publications

Identificación de un gen codificante de polifenol oxidasa (PPO) en Theobroma cacao L. (cacao) de Ecuador

Identificación de un gen codificante de polifenol oxidasa (PPO) en Theobroma cacao L. (cacao) de Ecuador

Analysis of gene expression patterns and levels in maize hybrids and their parents

Cloning and characterization of thioredoxin h in the three-pistil line of common wheat

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